Stroke is the third leading cause of death and affects nearly 1 million people in the U.S. each year. The cost for rehabilitation of stroke survivors is over $40 billion per year in the U.S. The debilitating effects of stroke derive from the loss of brain cells following the ischemic insult. Brain cells do not die instantly, but rather expire over several hours. This death of brain cells can be prevented by neurotrophic factors, a process called neuroprotection. If neurotrophins are injected directly into the brain within 3 hours of the stroke, then over half of affected brain cells can be saved. There presently is no neuroprotective agent available for stroke patients, because the neurotrophins, and other potential neuroprotective agents, do not cross the blood-brain barrier (BBB). It is unlikely that the BBB problem will be soon solved, because no pharmaceutical company in the U.S. has a BBB drug-targeting program. The Company's expertise is in the development of BBB drug targeting technologies, and has genetically engineered a molecular Trojan horse that crosses the human BBB via a specific receptor-transport system. In the present work, a neurotrophic factor will be fused to the molecular Trojan horse with genetic engineering for production of a novel recombinant fusion protein. This fusion protein will be a bi-functional molecule, and will bind both (a) the BBB receptor/transport, to enable receptor-mediated transport into brain following intravenous administration, and (b) the cognate receptor of the neurotrophic factor on brain cells, to induce neuroprotection. The goals of this phase I SBIR grant are to (i) produce a novel expression plasmid that produces the fusion protein, (ii) express the fusion protein in cells, and (iii) demonstrate retention of bifunctionality of the fusion protein with receptor-specific assays. This work will provide the basis for studies in phase 2, which will enable large-scale production of the fusion protein for Pharmacology/Toxicology studies required for an IND for the acute treatment of human stroke. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]